timing of immunophenotyping staining

From: maciej simm (flowlab@myflow.net)
Date: Fri Jun 28 2002 - 12:52:56 EST


Hello everyone,

I've asked a student to perform analysis of CD69 up-regulation (whole blood
+CD2/CD2r complex) over the course of first 4 hours with 15 minute snapshots
(16 tubes total). I am wondering if we should put the tubes in the fridge or
mix them with azide so we could stain all 16 at the end, or,  stain/lyse
right away? If anyone has done similar assays I would appreciate a comment.
It would be a lot easier to just stain everything in one shot.

The second issue is - do we start with one tube with 2 mL blood from which
we would take out a little bit every 15 min or start with 16 separate
smaller volume tubes? Someone commented that taking blood from the
single-tube-setup would change the environment for the remaining cells and
thus we would get a different result.

Maciej
--
Maciej S. Simm
Blackbelt Champion of Pipetting in the 2-20uL Range
Weill Cornell Medical College
Pediatrics Hematology/Oncology
Immunology Laboratory/Flow Cytometry Unit
525 East 68th Street N-805
New York, NY 10021
voice (212)746 3428
fax (212)746 8573
--



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